Cilia pp 169-192 | Cite as

STED and STORM Superresolution Imaging of Primary Cilia

  • T. Tony Yang
  • Weng Man Chong
  • Jung-Chi LiaoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1454)


The characteristic lengths of molecular arrangement in primary cilia are below the diffraction limit of light, challenging structural and functional studies of ciliary proteins. Superresolution microscopy can reach up to a 20 nm resolution, significantly improving the ability to map molecules in primary cilia. Here we describe detailed experimental procedure of STED microscopy imaging and dSTORM imaging, two of the most powerful superresolution imaging techniques. Specifically, we emphasize the use of these two methods on imaging proteins in primary cilia.

Key words

Primary cilium Ciliary protein Superresolution microscopy Diffraction limit Fluorophore STED STORM 



Bleaching/blinking assisted localization microscopy (BaLM)


Continuous wave (CW)


Direct stochastic optical reconstruction microscopy (dSTORM)


Fluorescence photoactivation localization microscopy (FPALM)


Ground state depletion (GSD)


Ground state depletion microscopy followed by individual molecule return (GSDIM)


Gated stimulated emission depletion (gSTED)


Intraflagellar transport (IFT)


Point accumulation for imaging in nanoscale topography (PAINT)


Photoactivated localization microscopy (PALM)


PALM with independently running acquisition (PALMIRA)


Point spread function (PSF)


Reversible saturable optical fluorescence transitions (RESOLFT)


Structured illumination microscopy (SIM)


Superresolution optical fluctuation imaging (SOFI)


Saturated structured illumination microscopy (SSIM)


Stimulated emission depletion (STED)


Stochastic optical reconstruction microscopy (STORM)



This work was supported by the Ministry of Science and Technology, Taiwan (Grant No. 103-2112-M-001-039-MY3), Academia Sinica Career Development Award, and Academia Sinica Nano Program.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute of Atomic and Molecular SciencesAcademia SinicaTaipeiTaiwan

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